超声波辅助法提取三叶青原花青素工艺的研究
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摘要
以三叶青块根为原料提取原花青素。通过使用超声波辅助法提取原花青素,以葡萄籽原花青素作为分析标品,利用香草醛-盐酸法绘制原花青素含量标准曲线,并对三叶青原花青素含量进行初步定量分析。单因素提取试验分析结果表明,乙醇体积分数、料液比、浸提时间和浸提温度4因素对三叶青原花青素的提取率影响较大,可作为后续工艺优化的主要控制因素。正交试验分析结果显示了最优提取工艺条件为乙醇体积分数为50%,料液比为1∶20,浸提温度为40℃,浸提时间为20 min,此条件下的原花青素提取率可达最高。
An ultrasonic-assisted extraction of proanthocyanidins from Tetrastigma hemsleyanum rootage was carried out optimized by using orthogonal experiment.Grape seed Proanthocyanidins was used as authentic standard to work out the calibration curve with vanillin-HCl assay method to measure the proanthocyanidin in the samples.The results of single factor extraction showed that the effects of ethanol volume fraction,solid-liquid ratio,extraction time and extraction temperature on the extraction rate of proanthocyanidins were significant,which could be used as the main factors for subsequent process optimization.The orthogonal experiment showed that the optimal extraction conditions were as follows:an ethanol concentration of 50%,a ratio of liquid to material of 1∶20(g/mL),a temperature of 40℃ and an extraction time of 20 min,by which the yield of proanthocyanidins was the highest.
An ultrasonic-assisted extraction of proanthocyanidins from Tetrastigma hemsleyanum rootage was carried out optimized by using orthogonal experiment.Grape seed Proanthocyanidins was used as authentic standard to work out the calibration curve with vanillin-HCl assay method to measure the proanthocyanidin in the samples.The results of single factor extraction showed that the effects of ethanol volume fraction,solid-liquid ratio,extraction time and extraction temperature on the extraction rate of proanthocyanidins were significant,which could be used as the main factors for subsequent process optimization.The orthogonal experiment showed that the optimal extraction conditions were as follows:an ethanol concentration of 50%,a ratio of liquid to material of 1∶20(g/mL),a temperature of 40℃ and an extraction time of 20 min,by which the yield of proanthocyanidins was the highest.
引文
[1] 李瑛琦,陆文超,于治国.三叶青的化学成分研究[J].中草药,2003,34(11):982-983.LI Y Q,LU W C,YU Z G.Research on Tetrastigma hemsleyanum diels et gilg chemical composition[J].Chinese Traditional and Herbal Drugs,2003,34(11):982-983.(in Chinese)
[2] 陈丽芸,郭素华.三叶青的化学成分及药理作用研究进展[J].浙江中医药大学学报,2012,36(12):1368-1370.
[3] 李鑫,刘景玲,李彦,等.三叶青原花青素结构分析[J].食品科学,2018,39(16):247-255.LI X,LIU J L,LI Y,et al.Structural analysis of procyanidins from Tetrastigma hemsleyanum diels et gilg[J].Food Science,2018,39(16):247-255.(in Chinese)
[4] FRACASSETTI D,COSTA C,MOULAY L,et al.Ellagic acid derivatives,ellagitannins,proanthocyanidins and other phenolics,vitamin C and antioxidant capacity of two powder products from camu-camu fruit (Myrciaria dubia)[J].Food Chemistry,2013,139(1/4):578-588.
[5] LIU Z Q,YANG Q L,ZHANG C S,et al.Study on antioxidant activity of proanthocyanidins from peanut skin[J].Advanced Materials Research,2011,197-198:1582-1586.
[6] TAICHI N,YOSHIKO F,YUKO S,et al.Procyanidins are potent inhibitors of LOX-1:a new player in the french paradox[J].Proceedings of the Japan Academy,2011,87(3):104.
[7] HOLT R R,HEISS C,KELM M,et al.The potential of flavanol and procyanidin intake to influence age-related vascular disease[J].Journal of Nutrition in Gerontology & Geriatrics,2012,31(3):290-323.
[8] YAMAKOSHI J,KATAOKA S,KOGA T,et al.Proanthocyanidin-rich extract from grape seeds attenuates the development of aortic atherosclerosis in cholesterol-fed rabbits[J].Atherosclerosis,1999,142(1):139-149.
[9] SANO A,UCHIDA R,SAITO M,et al.Beneficial effects of grape seed extract on malondialdehyde-modified LDL[J].Journal of Nutritional Science & Vitaminology,2007,53(2):174-182.
[10] 马亚兵,高海青,伊永亮,等.葡萄籽原花青素降低动脉粥样硬化兔血清C反应蛋白水平[J].中国动脉硬化杂志,2004,12(5):549-552.MA Y B,GAO H Q,YI Y L,et al.The serum c-reactive protein level of atherosclerotic rabbits was decreased by procyanidins of grape seed [J].Chinese Journal of Arteriosclerosis ,2004,12(5):549-552.(in Chinese)
[11] ZHAO G,GAO H,JIE Q,et al.The molecular mechanism of protective effects of grape seed proanthocyanidin extract on reperfusion arrhythmias in rats in vivo[J].Biological & Pharmaceutical Bulletin,2010,33(5):759-767.
[12] SHAO Z H,WOJCIK K R,DOSSUMBEKOVA A,et al.Grape seed proanthocyanidins protect cardiomyocytes from ischemia and reperfusion injury via Akt-NOS signaling[J].Journal of Cellular Biochemistry,2009,107(4):697-705.
[13] BELCARO G,LEDDA A,HU S,et al.Grape seed procyanidins in pre-and mild hypertension:a registry study[J].Evidence-Based Complementary and Alternative Mmedicine,2013,2013(6):1-5.
[14] IDOWU T O,OGUNDAINI A O,SALAU A O,et al.Doubly linked,A-type proanthocyanidin trimer and other constituents of Ixora coccinea leaves and their antioxidant and antibacterial properties[J].Phytochemistry,2010,71(17/18):2092.
[15] AL-HABIB A,AL-SALEH E,SAFER A M,et al.Bactericidal effect of grape seed extract on methicillin-resistant Staphylococcus aureus (MRSA)[J].Journal of Toxicological Sciences,2010,35(3):357-364.
[16] VERSTRAETEN S V,JAGGERS G K,FRAGA C G,et al.Procyanidins can interact with Caco-2 cell membrane lipid rafts:involvement of cholesterol[J].Biochimica et Biophysica Acta (BBA) - Biomembranes,2013,1828(11):2646-2653.
[17] ZHANG H,YERIGUI,YANG Y,et al.Structures and antioxidant and intestinal disaccharidase inhibitory activities of A-type proanthocyanidins from peanut skin[J].Journal of Agricultural & Food Chemistry,2013,61(37):8814.
[18] CEDó L,CASTELL-AUVí A,PALLARèS V,et al.Grape seed procyanidin extract modulates proliferation and apoptosis of pancreatic beta-cells[J].Food Chemistry,2013,138(1):524-530.
[19] CASTELLAUVí A,CEDó L,PALLARèS V,et al.Procyanidins modify insulinemia by affecting insulin production and degradation[J].Journal of Nutritional Biochemistry,2012,23(12):1565.
[20] 杨滢滢,王雪青,庞广昌.原花青素抗肿瘤作用机制研究进展[J].食品科学,2008,29(10):694-697.YANG Y Y,WANG X Q,PANG G C.Advances in research on the mechanism of procyanidins antitumor action[J].Food Science,2008,29(10):694-697.(in Chinese)
[21] CONNOR C A,ADRIAENS M,PIERINI R,et al.Procyanidin induces apoptosis of esophageal adenocarcinoma cells via JNK activation of c-Jun[J].Nutrition & Cancer,2014,66(2):335-341.
[22] SZEWCZYK K.Procyanidins from evening primrose,defatted seeds inhibit Invasiveness of breast cancer cells and modulate the expression of selected genes involved in angiogenesis,metastasis,and apoptosis[J].Nutrition & Cancer,2013,65(8):1219-1231.
[23] 李佳桥,余修亮,曾林晖,等.响应面试验优化超声波辅助提取莲房原花青素工艺[J].食品科学,2016,37(12):40-45.LI J Q,YU X L,ZENG L H,et al.The response surface test optimizes the ultrasonic assisted extraction process of lotus room procyanidins[J].Food Science,2016,37(12):40-45.(in Chinese)
[24] 马烨.红米原花青素的提取纯化与抗氧化活性研究[D].南昌:南昌大学,2016.
[25] 韦琴,乐薇,吕凯波,等.板栗壳原花青素的提取及体外抑菌作用研究[J].食品与发酵工业,2016,42(4):214-219.WEI Q,LE W,LYU K B,et al.Study on extraction of proanthocyanin from chestnut shell and its bacteriostasis in vitro[J].Food and Fermentation Industries,2016,42(4):214-219.(in Chinese)
[26] 李春阳,许时婴,王璋.香草醛-盐酸法测定葡萄籽、梗中原花青素含量的研究[J].食品工业科技,2004,25(6):157-161.LI C Y,XU S Y,WANG Z.Determination of procyanidins in grape seed and stem by vanillin-hydrochloric acid method[J].Science and Technology of Food Industry,2004,25(6):157-161.(in Chinese)
[27] 周秋枝,黄蕾,沈丹华,等.火棘果中原花青素含量测定方法的建立[J].食品工业科技,2013,34(7):314-318.ZHOU Q Z,HUANG L,SHEN D H,et al.Establishment of a method for the determination of procyanidins in echinoderm[J].Science and Technology of Food Industry,2013,34(7):314-318.(in Chinese)
[28] 尤新军,郭蕊,王琳,等.黄精总皂苷超声提取工艺研究[J].西北林学院学报,2010,25(3):163-166.YOU X J,GUO R,WANG L,et al.Ultrasonic extraction process of saponins from Polygonatum sibiricun[J].Journal of Northwest Forestry University,2010,25(3):163-166.(in Chinese)
[29] 刘婷,金瑞.超声波法提取西洋参多糖的工艺研究[J].西北林学院学报,2014,29(3):174-177.LIU T,JIN R.Technology of ultrasonic wave assisted extraction of polysaccharides from Panax quinque folium[J].Journal of Northwest Forestry University,2014,29(3):174-177.(in Chinese)
[30] 黄尚荣,杨雪娜,张露,等.龙眼皮原花青素提取工艺优化及其抗氧化活性测定[J].食品科学,2014,35(10):68-75.HUANG S R,YANG X N,ZHANG L,et al.Optimization of extraction process and determination of antioxidant activity of procyanidins from longeyelid[J].Food Science,2014,35(10):68-75.(in Chinese)
[31] 任小娜,陈志梅,曾俊,等.黑果枸杞中原花青素提取条件的优化与含量测定[J].食品与发酵工业,2015,41(1):147-150.REN X N,CHEN Z M,ZENG J,et al.Optimization of extraction conditions and content determination of procyanidins from wolfberry[J].Food and Fermentation Industries,2015,41(1):147-150.(in Chinese)
[32] 杨玲,刘虹男,张冬严,等.紫叶白桦组培苗叶片花青素含量影响因素分析[J].森林工程,2017,33(2):33-40.YANG L,LIU H N,ZHANG Y D,et al.Analysis of factors influencing on anthocyanin content in leaves of tissue culture plantlet of Royal Frost Betula[J].Forest Engineering,2017,33(2):33-40.(in Chinese)
[33] 张镜,何洪洛.正交试验优化大叶榕果实原花青素提取工艺[J].食品科学,2015,36(2):41-45.ZHANG J,HE H L.Orthogonal experiment was conducted to optimize extraction technology of procyanidins from ficus macrophylla fruit[J].Food Science,2015,36(2):41-45.(in Chinese)
[34] 秦超,容蓉,杨勇,等.多指标正交试验优选麻黄附子细辛汤提取工艺[J].中国实验方剂学杂志,2012,18(9):35-39.QIN C,RONG R,YANG Y,et al.The extraction technology of asarum cassia decoction from ephedra rhizome was optimized by multi-index orthogonal test[J].Chinese Journal of Experimental Traditional Medical Formulae,2012,18(9):35-39.(in Chinese)
[2] 陈丽芸,郭素华.三叶青的化学成分及药理作用研究进展[J].浙江中医药大学学报,2012,36(12):1368-1370.
[3] 李鑫,刘景玲,李彦,等.三叶青原花青素结构分析[J].食品科学,2018,39(16):247-255.LI X,LIU J L,LI Y,et al.Structural analysis of procyanidins from Tetrastigma hemsleyanum diels et gilg[J].Food Science,2018,39(16):247-255.(in Chinese)
[4] FRACASSETTI D,COSTA C,MOULAY L,et al.Ellagic acid derivatives,ellagitannins,proanthocyanidins and other phenolics,vitamin C and antioxidant capacity of two powder products from camu-camu fruit (Myrciaria dubia)[J].Food Chemistry,2013,139(1/4):578-588.
[5] LIU Z Q,YANG Q L,ZHANG C S,et al.Study on antioxidant activity of proanthocyanidins from peanut skin[J].Advanced Materials Research,2011,197-198:1582-1586.
[6] TAICHI N,YOSHIKO F,YUKO S,et al.Procyanidins are potent inhibitors of LOX-1:a new player in the french paradox[J].Proceedings of the Japan Academy,2011,87(3):104.
[7] HOLT R R,HEISS C,KELM M,et al.The potential of flavanol and procyanidin intake to influence age-related vascular disease[J].Journal of Nutrition in Gerontology & Geriatrics,2012,31(3):290-323.
[8] YAMAKOSHI J,KATAOKA S,KOGA T,et al.Proanthocyanidin-rich extract from grape seeds attenuates the development of aortic atherosclerosis in cholesterol-fed rabbits[J].Atherosclerosis,1999,142(1):139-149.
[9] SANO A,UCHIDA R,SAITO M,et al.Beneficial effects of grape seed extract on malondialdehyde-modified LDL[J].Journal of Nutritional Science & Vitaminology,2007,53(2):174-182.
[10] 马亚兵,高海青,伊永亮,等.葡萄籽原花青素降低动脉粥样硬化兔血清C反应蛋白水平[J].中国动脉硬化杂志,2004,12(5):549-552.MA Y B,GAO H Q,YI Y L,et al.The serum c-reactive protein level of atherosclerotic rabbits was decreased by procyanidins of grape seed [J].Chinese Journal of Arteriosclerosis ,2004,12(5):549-552.(in Chinese)
[11] ZHAO G,GAO H,JIE Q,et al.The molecular mechanism of protective effects of grape seed proanthocyanidin extract on reperfusion arrhythmias in rats in vivo[J].Biological & Pharmaceutical Bulletin,2010,33(5):759-767.
[12] SHAO Z H,WOJCIK K R,DOSSUMBEKOVA A,et al.Grape seed proanthocyanidins protect cardiomyocytes from ischemia and reperfusion injury via Akt-NOS signaling[J].Journal of Cellular Biochemistry,2009,107(4):697-705.
[13] BELCARO G,LEDDA A,HU S,et al.Grape seed procyanidins in pre-and mild hypertension:a registry study[J].Evidence-Based Complementary and Alternative Mmedicine,2013,2013(6):1-5.
[14] IDOWU T O,OGUNDAINI A O,SALAU A O,et al.Doubly linked,A-type proanthocyanidin trimer and other constituents of Ixora coccinea leaves and their antioxidant and antibacterial properties[J].Phytochemistry,2010,71(17/18):2092.
[15] AL-HABIB A,AL-SALEH E,SAFER A M,et al.Bactericidal effect of grape seed extract on methicillin-resistant Staphylococcus aureus (MRSA)[J].Journal of Toxicological Sciences,2010,35(3):357-364.
[16] VERSTRAETEN S V,JAGGERS G K,FRAGA C G,et al.Procyanidins can interact with Caco-2 cell membrane lipid rafts:involvement of cholesterol[J].Biochimica et Biophysica Acta (BBA) - Biomembranes,2013,1828(11):2646-2653.
[17] ZHANG H,YERIGUI,YANG Y,et al.Structures and antioxidant and intestinal disaccharidase inhibitory activities of A-type proanthocyanidins from peanut skin[J].Journal of Agricultural & Food Chemistry,2013,61(37):8814.
[18] CEDó L,CASTELL-AUVí A,PALLARèS V,et al.Grape seed procyanidin extract modulates proliferation and apoptosis of pancreatic beta-cells[J].Food Chemistry,2013,138(1):524-530.
[19] CASTELLAUVí A,CEDó L,PALLARèS V,et al.Procyanidins modify insulinemia by affecting insulin production and degradation[J].Journal of Nutritional Biochemistry,2012,23(12):1565.
[20] 杨滢滢,王雪青,庞广昌.原花青素抗肿瘤作用机制研究进展[J].食品科学,2008,29(10):694-697.YANG Y Y,WANG X Q,PANG G C.Advances in research on the mechanism of procyanidins antitumor action[J].Food Science,2008,29(10):694-697.(in Chinese)
[21] CONNOR C A,ADRIAENS M,PIERINI R,et al.Procyanidin induces apoptosis of esophageal adenocarcinoma cells via JNK activation of c-Jun[J].Nutrition & Cancer,2014,66(2):335-341.
[22] SZEWCZYK K.Procyanidins from evening primrose,defatted seeds inhibit Invasiveness of breast cancer cells and modulate the expression of selected genes involved in angiogenesis,metastasis,and apoptosis[J].Nutrition & Cancer,2013,65(8):1219-1231.
[23] 李佳桥,余修亮,曾林晖,等.响应面试验优化超声波辅助提取莲房原花青素工艺[J].食品科学,2016,37(12):40-45.LI J Q,YU X L,ZENG L H,et al.The response surface test optimizes the ultrasonic assisted extraction process of lotus room procyanidins[J].Food Science,2016,37(12):40-45.(in Chinese)
[24] 马烨.红米原花青素的提取纯化与抗氧化活性研究[D].南昌:南昌大学,2016.
[25] 韦琴,乐薇,吕凯波,等.板栗壳原花青素的提取及体外抑菌作用研究[J].食品与发酵工业,2016,42(4):214-219.WEI Q,LE W,LYU K B,et al.Study on extraction of proanthocyanin from chestnut shell and its bacteriostasis in vitro[J].Food and Fermentation Industries,2016,42(4):214-219.(in Chinese)
[26] 李春阳,许时婴,王璋.香草醛-盐酸法测定葡萄籽、梗中原花青素含量的研究[J].食品工业科技,2004,25(6):157-161.LI C Y,XU S Y,WANG Z.Determination of procyanidins in grape seed and stem by vanillin-hydrochloric acid method[J].Science and Technology of Food Industry,2004,25(6):157-161.(in Chinese)
[27] 周秋枝,黄蕾,沈丹华,等.火棘果中原花青素含量测定方法的建立[J].食品工业科技,2013,34(7):314-318.ZHOU Q Z,HUANG L,SHEN D H,et al.Establishment of a method for the determination of procyanidins in echinoderm[J].Science and Technology of Food Industry,2013,34(7):314-318.(in Chinese)
[28] 尤新军,郭蕊,王琳,等.黄精总皂苷超声提取工艺研究[J].西北林学院学报,2010,25(3):163-166.YOU X J,GUO R,WANG L,et al.Ultrasonic extraction process of saponins from Polygonatum sibiricun[J].Journal of Northwest Forestry University,2010,25(3):163-166.(in Chinese)
[29] 刘婷,金瑞.超声波法提取西洋参多糖的工艺研究[J].西北林学院学报,2014,29(3):174-177.LIU T,JIN R.Technology of ultrasonic wave assisted extraction of polysaccharides from Panax quinque folium[J].Journal of Northwest Forestry University,2014,29(3):174-177.(in Chinese)
[30] 黄尚荣,杨雪娜,张露,等.龙眼皮原花青素提取工艺优化及其抗氧化活性测定[J].食品科学,2014,35(10):68-75.HUANG S R,YANG X N,ZHANG L,et al.Optimization of extraction process and determination of antioxidant activity of procyanidins from longeyelid[J].Food Science,2014,35(10):68-75.(in Chinese)
[31] 任小娜,陈志梅,曾俊,等.黑果枸杞中原花青素提取条件的优化与含量测定[J].食品与发酵工业,2015,41(1):147-150.REN X N,CHEN Z M,ZENG J,et al.Optimization of extraction conditions and content determination of procyanidins from wolfberry[J].Food and Fermentation Industries,2015,41(1):147-150.(in Chinese)
[32] 杨玲,刘虹男,张冬严,等.紫叶白桦组培苗叶片花青素含量影响因素分析[J].森林工程,2017,33(2):33-40.YANG L,LIU H N,ZHANG Y D,et al.Analysis of factors influencing on anthocyanin content in leaves of tissue culture plantlet of Royal Frost Betula[J].Forest Engineering,2017,33(2):33-40.(in Chinese)
[33] 张镜,何洪洛.正交试验优化大叶榕果实原花青素提取工艺[J].食品科学,2015,36(2):41-45.ZHANG J,HE H L.Orthogonal experiment was conducted to optimize extraction technology of procyanidins from ficus macrophylla fruit[J].Food Science,2015,36(2):41-45.(in Chinese)
[34] 秦超,容蓉,杨勇,等.多指标正交试验优选麻黄附子细辛汤提取工艺[J].中国实验方剂学杂志,2012,18(9):35-39.QIN C,RONG R,YANG Y,et al.The extraction technology of asarum cassia decoction from ephedra rhizome was optimized by multi-index orthogonal test[J].Chinese Journal of Experimental Traditional Medical Formulae,2012,18(9):35-39.(in Chinese)